Blockers of G-protein coupled receptors (GPCRs), angiotensin II (Ang II) type 1 (AT(1)) receptor and beta(1)-adrenergic (Ad) receptor, have been shown to improve the prognosis of cardiovascular disease. Cholesterol molecules in the cell membrane are needed to stabilize GPCRs as well as the cell membrane itself. We determined whether the functions of AT(1) and beta(1)-Ad receptors were changed by cholesterol depletion from cardiovascular cell membranes. Ang II-induced inositol phosphate production through AT(1) receptor was suppressed by cholesterol depletion from cell membranes using rosuvastatin or methyl-beta-cyclodextrin (M beta CD), whereas isoproterenol-induced cyclic AMP production through beta(1)-Ad receptor did not change after cholesterol depletion. In addition, the binding affinities of Ang II and AT(1) receptor blocker after cholesterol depletion were significantly lower than those before depletion. Although AT(1) receptor expression levels did not change after cholesterol depletion, the expression levels of AT(1) receptor that could bind to Ang II significantly decreased after depletion. The changes in the structure of AT(1) receptor due to depletion were confirmed by substituted-cysteine accessibility mapping. In conclusion, Ang II-induced activation of AT(1) receptor is reduced without affecting the function of beta(1)-Ad receptor after cholesterol depletion from cardiovascular cell membranes. (C) 2019 Elsevier Inc. All rights reserved.